Case of the Rearranging Plasmid?

[Graham Dellaire]

I have prepared a plasmid based vector that contains a large terminal fragment of
the adenovirus-2 genome.  The vector is 22 kbp and has the odd and frustrating
ability to recombine and rearrange when grown in bacteria in large culture volumes
(greater than 5 ml!!)  When I grow the bacteria (Dh5 or10) in 5 ml cultures I get no
rearrangment but as soon as I try to grow a larger culture volume I end up with
a ladder of plasmids or a new one all together!!!

Is it perhaps a problem with adeno DNA in bacteria... perhaps expression of one
of the adeno genes etc?

ANyone have similar problems? and most importantly what was your solution???<g>



Thanks in advance for you time and help

Graham
Mcgill, Exp Med.

popa0206 at po-box.mcgill.ca